Method for detecting DNA basic group mutation

Method for detecting DNA basic group mutation

  • CN 101,838,688 B
  • Filed: 04/07/2008
  • Issued: 03/12/2014
  • Est. Priority Date: 04/07/2008
  • Status: Active Grant
First Claim
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1. detect a method that whether has base mutation in DNA, comprise the following steps successively:

  • 1) prepare strand polynucleotide K1, the normal sequence complete complementary of described K1 and DNA to be measured;

    2) prepare strand polynucleotide K2, the middle portion of described K2 has DNAzyme activity, and K2 and K1 have 15nt complete complementary at least, and includes at least DNAzyme sequence of 10nt, and K2 and K1 at least respectively have 6nt not complementary end to end;

    3) prepare DNAzyme substrate S, use fluorescein-labelled S;

    DNA part and the K2 complete complementary of substrate S, have the restriction enzyme site that comprises two special RNA bases in the middle of substrate S;

    4) K1 and K2 are mixed, then add substrate S, the compound that adds DNA to be measured and following formula (I), the control systems that the compound add DNA fragmentation that the normal sequence by DNA to be measured forms and following formula (I) is set simultaneously, whether definite DNA to be measured there is sudden change as follows;

    if the I of DNA reaction system to be measured 424nm/ I 527nmbe less than the I of control systems 424nm/ I 527nm, there is base mutation in DNA to be measured;

    Described I 424nmfluorescence intensity for the emission wavelength emission peak that is 424nm;

    Described I 527nmfluorescence intensity for the emission wavelength emission peak that is 527nm.

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